Abstract
Main conclusion
We observed a close correlation between the inhibition of photosystem II and the oxidation of polyphenols during an acute oxidative stress in sunflower leaf discs.
Abstract
To assess the physiological significance of polyphenols as antioxidants in planta, we compared the kinetics of polyphenols oxidation with the inhibition of the photosynthetic apparatus in sunflower leaf discs exposed to an acute photooxidative stress. Illumination of leaf discs in the presence of methyl viologen induced a rapid and large non-photochemical quenching of chlorophyll-a fluorescence, which was reversed after 4 h of treatment as indicated by the ≈ 30% increases of the steady-state (Fs) and maximal (Fm′) levels of chlorophyll-a fluorescence relative to the first hour of treatment. This event coincided with the accelerated decreases of the maximum (Fv/Fm) and effective (∆F/Fm′) quantum yields of photosystem II, and also with the beginning of polyphenols oxidation, estimated by the UV absorbance of methanolic leaf extracts, and supported by the Folin–Ciocalteu method and cyclic voltammetry. The decreases of Fv/Fm and the concentrations of reducing polyphenols were highly correlated (R2 = 0.877) during the experiment. Coherent with the decrease of UV absorbance of methanolic extracts, polyphenol oxidation resulted in a marked decrease of UV absorbance of leaf epidermis. Also, polymerization of oxidized polyphenols caused the accumulation of brown pigments in the MV-treated leaf discs, decreasing leaf reflectance, especially at 550 and 740 nm. Fluorescence intensities were also decreased during the MV treatment. Interestingly, the emission fluorescence ratio F740/F684 (excitation at 550 nm) decreased similarly to Fv/Fm (R2 = 0.981) due to the brown pigments. Moreover, the excitation fluorescence ratio F484/F680 (emission at 740 nm) was linearly correlated (R2 = 0.957) to ∆F/Fm′, indicating a decrease of efficiency of energy transfer between the antenna pigments to the photosystem II reaction center during the oxidative stress. These results support the view that polyphenols can be effective antioxidants protecting the plants against reactive oxygen species.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agati G, Mazzinghi P, Fusi F, Ambrosini I (1995) The F685/F730 chlorophyll fluorescence ratio as a tool in plant physiology: response to physiological and environmental factors. J Plant Physiol 145(3):228–238. https://doi.org/10.1016/S0176-1617(11)81882-1
Agati G, Mazzinghi P, Lipucci Di Paola M, Fusi F, Cecchi G (1996) The F685/F730 chlorophyll fluorescence ratio as indicator of chilling stress in plants. J Plant Physiol 148(3–4):384–390. https://doi.org/10.1016/S0176-1617(96)80270-7
Agati G, Matteini P, Goti A, Tattini M (2007) Chloroplast-located flavonoids can scavenge singlet oxygen. New Phytol 174(1):77–89. https://doi.org/10.1111/j.1469-8137.2007.01986.x
Agati G, Azzarello E, Pollastri S, Tattini M (2012) Flavonoids as antioxidants in plants: location and functional significance. Plant Sci 196:67–76. https://doi.org/10.1016/j.plantsci.2012.07.014
Bertolini A, Petrussa E, Patui S, Zancani M, Peresson C, Casolo V, Vianello A, Braidot E (2016) Flavonoids and darkness lower PCD in senescing Vitis vinifera suspension cell cultures. BMC Plant Biol 16(1):233. https://doi.org/10.1186/s12870-016-0917-y
Buschmann C, Lichtenthaler HK (1998) Principles and characteristics of multi-colour fluorescence imaging of plants. J Plant Physiol 152(2–3):297–314. https://doi.org/10.1016/S0176-1617(98)80144-2
Cerovic ZG, Ounis A, Cartelat A, Latouche G, Goulas Y, Meyer S, Moya I (2002) The use of chlorophyll fluorescence excitation spectra for the non-destructive in situ assessment of UV-absorbing compounds in leaves. Plant Cell Environ 25(12):1663–1676. https://doi.org/10.1046/j.1365-3040.2002.00942.x
Derks A, Schaven K, Bruce D (2015) Diverse mechanisms for photoprotection in photosynthesis. Dynamic regulation of photosystem II excitation in response to rapid environmental change. Biochim Biophys Acta Bioenerg 1847(4–5):468–485. https://doi.org/10.1016/j.bbabio.2015.02.008
Dixon RA, Paiva NL (1995) Stress-induced phenylpropanoid metabolism. Plant Cell 7(7):1085–1097. https://doi.org/10.1105/tpc.7.7.1085
Dong J, Wan G, Liang Z (2010) Accumulation of salicylic acid-induced phenolic compounds and raised activities of secondary metabolic and antioxidative enzymes in Salvia miltiorrhiza cell culture. J Biotechnol 148(2–3):99–104. https://doi.org/10.1016/j.jbiotec.2010.05.009
Edreva A (2005) The importance of non-photosynthetic pigments and cinnamic acid derivatives in photoprotection. Agric Ecosyst Environ 106:135–146. https://doi.org/10.1016/j.agee.2004.10.002
Ghasemzadeh A, Ghasemzadeh N (2011) Flavonoids and phenolic acids: role and biochemical activity in plants and human. J Med Plant Res 5(31):6697–6703. https://doi.org/10.5897/JMPR11.1404
Gitelson AA, Buschmann C, Lichtenthaler HK (1999) The chlorophyll fluorescence ratio F735/F700 as an accurate measure of the chlorophyll content in plants. Remote Sens Environ 69(3):296–302. https://doi.org/10.1016/S0034-4257(99)00023-1
Hayat Q, Hayat S, Irfan M, Ahmad A (2010) Effect of exogenous salicylic acid under changing environment: a review. Environ Exp Bot 68(1):14–25. https://doi.org/10.1016/j.envexpbot.2009.08.005
Hernández I, Alegre L, Munné-Bosch S (2006) Enhanced oxidation of flavan-3-ols and proanthocyanidin accumulation in water-stressed tea plants. Phytochemistry 67(11):1120–1126. https://doi.org/10.1016/j.phytochem.2006.04.002
Hernández I, Alegre L, Van Breusegem F, Munné-Bosch S (2009) How relevant are flavonoids as antioxidants in plants? Trends Plant Sci 14(3):125–132. https://doi.org/10.1016/j.tplants.2008.12.003
Hernanz-Vila D, Jara-Palacios MJ, Escudero-Gilete ML, Heredia FJ (2017) Applications of voltammetric analysis to wine products. In: Stoytcheva M, Zlatev R (eds) Applications of the voltammetry. IntechOpen Limited, London, pp 109–127. https://doi.org/10.5772/67696
Jaiswal S, Chawla R, Sawhney S (2012) Correlations among attributes of senescence and antioxidative status of leaf discs during epiphyllous bud differentiation in Kalanchoe pinnata Lam. (Pers.). Z Naturforsch Sect C J Biosci 67 C(7–8):418–428. https://doi.org/10.1515/znc-2012-7-810
Kilmartin PA, Hsu CF (2003) Characterisation of polyphenols in green, oolong, and black teas, and in coffee, using cyclic voltammetry. Food Chem 82(4):501–512. https://doi.org/10.1016/S0308-8146(03)00066-9
Kilmartin PA, Zou H, Waterhouse AL (2001) A cyclic voltammetry method suitable for characterizing antioxidant properties of wine and wine phenolics. J Agric Food Chem 49(4):1957–1965. https://doi.org/10.1021/jf001044u
Klughammer C, Schreiber U (2008) Complementary PS II quantum yields calculated from simple fluorescence parameters measured by PAM fluorometry and the Saturation Pulse method. PAM Appl Notes 1:27–35
Kováčik J, Grúz J, Bačkor M, Strnad M, Repčák M (2009) Salicylic acid-induced changes to growth and phenolic metabolism in Matricaria chamomilla plants. Plant Cell Rep 28(1):135–143. https://doi.org/10.1007/s00299-008-0627-5
Kováčik J, Klejdus B, Hedbavny J, Bačkor M (2010) Effect of copper and salicylic acid on phenolic metabolites and free amino acids in Scenedesmus quadricauda (Chlorophyceae). Plant Sci 178(3):307–311. https://doi.org/10.1016/j.plantsci.2010.01.009
Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382. https://doi.org/10.1016/0076-6879(87)48036-1
Louis J, Cerovic ZG, Moya I (2006) Quantitative study of fluorescence excitation and emission spectra of bean leaves. J Photochem Photobiol B Biol 85(1):65–71. https://doi.org/10.1016/j.jphotobiol.2006.03.009
Louis J, Meyer S, Maunoury-Danger F, Fresneau C, Meudec E, Cerovic ZG (2009) Seasonal changes in optically assessed epidermal phenolic compounds and chlorophyll contents in leaves of sessile oak (Quercus petraea): towards signatures of phenological stage. Funct Plant Biol 36(8):732–741. https://doi.org/10.1071/FP09010
Louis J, Genet H, Meyer S, Soudani K, Montpied P, Legout A, Dreyer E, Cerovic ZG, Dufrêne E (2012) Tree age-related effects on sun acclimated leaves in a chronosequence of beech (Fagus sylvatica) stands. Funct Plant Biol 39(4):323–331. https://doi.org/10.1071/FP11248
Mandal SM, Chakraborty D, Dey S (2010) Phenolic acids act as signaling molecules in plant-microbe symbioses. Plant Signal Behav 5(4):359–368. https://doi.org/10.4161/psb.5.4.10871
Meir S, Kanner J, Akiri B, Philosoph-Hadas S (1995) Determination and involvement of aqueous reducing compounds in oxidative defense systems of various senescing leaves. J Agric Food Chem 43(7):1813–1819. https://doi.org/10.1021/jf00055a012
Nakabayashi R, Saito K (2015) Integrated metabolomics for abiotic stress responses in plants. Curr Opin Plant Biol 24:10–16. https://doi.org/10.1016/j.pbi.2015.01.003
Nakabayashi R, Yonekura-Sakakibara K, Urano K, Suzuki M, Yamada Y, Nishizawa T, Matsuda F, Kojima M, Sakakibara H, Shinozaki K, Michael AJ, Tohge T, Yamazaki M, Saito K (2014) Enhancement of oxidative and drought tolerance in Arabidopsis by overaccumulation of antioxidant flavonoids. Plant J 77(3):367–379. https://doi.org/10.1111/tpj.12388
Neill SO, Gould KS, Kilmartin PA, Mitchell KA, Markham KR (2002) Antioxidant capacities of green and cyanic leaves in the sun species, Quintinia serrata. Funct Plant Biol 29(12):1437–1443. https://doi.org/10.1071/FP02100
Nichelmann L, Schulze M, Herppich WB, Bilger W (2016) A simple indicator for non-destructive estimation of the violaxanthin cycle pigment content in leaves. Photosynth Res 128(2):183–193. https://doi.org/10.1007/s11120-016-0218-1
Ounis A, Cerovic Z, Briantais J-M, Moya I (2001) Dual-excitation FLIDAR for the estimation of epidermal UV absorption in leaves and canopies. Remote Sens Environ 76:33–48. https://doi.org/10.1016/S0034-4257(00)00190-5
Pérez FJ, Villegas D, Mejia N (2002) Ascorbic acid and flavonoid-peroxidase reaction as a detoxifying system of H2O2 in grapevine leaves. Phytochemistry 60(6):573–580. https://doi.org/10.1016/S0031-9422(02)00146-2
Pourcel L, Routaboul JM, Cheynier V, Lepiniec L, Debeaujon I (2007) Flavonoid oxidation in plants: from biochemical properties to physiological functions. Trends Plant Sci 12(1):29–36. https://doi.org/10.1016/j.tplants.2006.11.006
Prior RL, Wu X, Schaich K (2005) Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric Food Chem 53(10):4290–4302. https://doi.org/10.1021/jf0502698
Ramešová Š, Sokolová R, Degano I (2015) The study of the oxidation of the natural flavonol fisetin confirmed quercetin oxidation mechanism. Electrochim Acta 182:544–549. https://doi.org/10.1016/j.electacta.2015.09.144
Sava VM, Galkin BN, Hong MY, Yang PC, Huang GS (2001) A novel melanin-like pigment derived from black tea leaves with immuno-stimulating activity. Food Res Int 34(4):337–343. https://doi.org/10.1016/S0963-9969(00)00173-3
Shao L, Shu Z, Peng CL, Lin ZF, Yang CW, Gu Q (2008) Enhanced sensitivity of Arabidopsis anthocyanin mutants to photooxidation: a study with fluorescence imaging. Funct Plant Biol 35(8):714–724. https://doi.org/10.1071/FP08069
Sokolová R, Ramešová S, Degano I, Hromadová M, Gál M, Abka J (2012) The oxidation of natural flavonoid quercetin. Chem Commun 48(28):3433–3435. https://doi.org/10.1039/c2cc18018a
Stelzner J, Roemhild R, Garibay-Hernández A, Harbaum-Piayda B, Mock HP, Bilger W (2019) Hydroxycinnamic acids in sunflower leaves serve as UV-A screening pigments. Photochem Photobiol Sci 18(7):1649–1659. https://doi.org/10.1039/c8pp00440d
Takahama U (2004) Oxidation of vacuolar and apoplastic phenolic substrates by peroxidase: physiological significance of the oxidation reactions. Phytochem Rev 3(1–2):207–219. https://doi.org/10.1023/B:PHYT.0000047805.08470.e3
Takahama U, Hirotsu M, Oniki T (1999) Age-dependent changes in levels of ascorbic acid and chlorogenic acid, and activities of peroxidase and superoxide dismutase in the apoplast of tobacco leaves: mechanism of the oxidation of chlorogenic acid in the apoplast. Plant Cell Physiol 40(7):716–724. https://doi.org/10.1093/oxfordjournals.pcp.a029598
Tomás-Barberán FA, Gil MI, Castañer M, Artés F, Saltveit ME (1997) Effect of selected browning inhibitors on phenolic metabolism in stem tissue of harvested lettuce. J Agric Food Chem 45(3):583–589. https://doi.org/10.1021/jf960478f
Treutter D (2005) Significance of flavonoids in plant resistance and enhancement of their biosynthesis. Plant Biol 7(6):581–591. https://doi.org/10.1055/s-2005-873009
Yamasaki H, Sakihama Y, Ikehara N (1997) Flavonoid-peroxidase reaction as a detoxification mechanism of plant cells against H2O2. Plant Physiol 115(4):1405–1412. https://doi.org/10.1104/pp.115.4.1405
Zeng L, Wang Y, Zhou J (2016) Spectral analysis on origination of the bands at 437 nm and 475.5 nm of chlorophyll fluorescence excitation spectrum in Arabidopsis chloroplasts. Lumin 31(3):769–774. https://doi.org/10.1002/bio.3022
Acknowledgements
We are grateful to Lionel Saunois and Amandine Dubois from the greenhouse facility for taking care of the sunflower plants, and to Gwendal Latouche for his technical assistance. GS acknowledges his sabbatical leave from the Université du Québec à Trois-Rivières.
Author information
Authors and Affiliations
Contributions
GS and ZGC conceived research, conducted most experiments and analyzed the data. WMAER and PM conducted experiments with voltammetry and analyzed the data. GS prepared the figures and wrote most of the manuscript. All authors reviewed and approved the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
ZGC declares a link to the company FORCE-A as one of the co-authors of the Multiplex patent that the company exploits. Other authors have no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
425_2019_3316_MOESM1_ESM.docx
Fig. 1S Polyphenol oxidation estimated from leaf methanolic extracts. Absorbance spectra of extracts from leaf treated or not with methyl viologen for different times (DOCX 56 kb)
Rights and permissions
About this article
Cite this article
Samson, G., Cerovic, Z.G., El Rouby, W.M.A. et al. Oxidation of polyphenols and inhibition of photosystem II under acute photooxidative stress. Planta 251, 16 (2020). https://doi.org/10.1007/s00425-019-03316-x
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00425-019-03316-x